LOCUS       sequence-338404-        5513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRE-controlled mAID-fusion sleeping beauty transposon vector with 
            selection marker (hygromycin).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5513)
  AUTHORS   Yeung TK, Lau HW, Ma HT, Poon RYC
  TITLE     One-step multiplex toolkit for efficient generation of conditional 
            gene silencing human cell lines.
  JOURNAL   Mol Biol Cell. 2021 May 12:mbcE21020051. doi: 
            10.1091/mbc.E21-02-0051.
  PUBMED    33979199
REFERENCE   2  (bases 1 to 5513)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Biol 
            Cell. 2021 May 12:mbcE21020051. doi: 10.1091/mbc.E21-02-0051."
COMMENT     Sequence Label: pUHD-SB-mAID/Hyg
FEATURES             Location/Qualifiers
     source          1..5513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(311..329,330..352,353..371,372..394,395..413,414..436,
                     437..455,456..478,479..497,498..520,521..539,540..562,
                     563..581)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        614..652
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     649..673
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             760..963
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     polyA_signal    1032..1153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1043..1062)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1097..1116
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             1885..2904
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     primer_bind     complement(2959..2976)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2965..3189
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3515..3532)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3686..4274)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3766..3785)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4445..5236,5237..5305))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5068..5087
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5306..5410)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cagttgaagt cggaagttta catacactta agttggagtc attaaaactc gtttttcaac
       61 tactccacaa atttcttgtt aacaaacaat agttttggca agtcagttag gacatctact
      121 ttgtgcatga cacaagtcat ttttccaaca attgtttaca gacagattat ttcacttata
      181 attcactgta tcacaattcc agtgggtcag aagtttacat acactaagtt cgactcctct
      241 gcagaatgcg gcgatgtttc ggtaaggggt ccgctatcta ggccctttcg tctttagtcg
      301 agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc
      361 agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga
      421 aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc
      481 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa
      541 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagctcg
      601 gtacccgggt cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt
      661 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagagaccg
      721 ggaccgatcc agcctccgcg ggctagcatg gccggagcaa aggagaaatc agcctgccct
      781 aaagaccccg caaagccccc cgcaaaagca caggtggtgg gatggccccc cgtgcgcagc
      841 taccggaaga acgtgatggt gtcctgccag aagagctccg gaggaccaga ggcagcagcc
      901 tttgtgaagg tgtctatgga tggcgcccct tacctgagaa agatcgacct gaggatgtat
      961 aaggcctcca tggctcgaga attcgagctc ggtacccggg gatccagaca tgataagatc
     1021 gatagacatg ataagataca ttgatgagtt tggacaaacc acaacaagaa tgcagtgaaa
     1081 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     1141 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     1201 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtacctaga cgagaccctg
     1261 tctcacaaaa taaagtaagc ccggactgag tgcggaaagg cgggcctggc gggtctggtc
     1321 tccccatgcg ggccaccaga ggccctgcag ccttcagtcg cttgaagggg taatggcgct
     1381 tccactcaca aacatggcgg acagagcgtg tgaacgagat gaacagcccc tcaaaaatat
     1441 ggccgccgag gctggacggc cgtgccccag cagcaccgcc tccgcgcccc acgtgatctc
     1501 tcgccgggca cagcgctgac cgcggaggtc caaccggaag aatgtccgga ttggacattc
     1561 ggaagagggc ccgccttccc tggggaatct ctgcgcacgc gcagaacgct tcgaccaatg
     1621 aaaacacagg aagccgtccg cgcaaccgcg ttgcgtcact tctgccgccc ctgtttcaag
     1681 gtatatagcc gtagacggaa cttcgccttt ctctcggcct tagcgccatt tttttgggtg
     1741 agtgtttttt ggttcctgcg ttgggattcc gtgtacaatc catagacatc tgacctcggc
     1801 acttagcatc atcacagcaa actaactgta gcctttctct ctttccctgt agaaacctct
     1861 gcactcgaca tggatagatc cggaaagcct gaactcaccg cgacgtctgt cgagaagttt
     1921 ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct
     1981 cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc
     2041 gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt
     2101 ccggaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat ctcccgccgt
     2161 gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg
     2221 gtcgcggagg ctatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc
     2281 ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt
     2341 gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc
     2401 gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc
     2461 gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc
     2521 attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc
     2581 tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg
     2641 gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc
     2701 tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac
     2761 gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg
     2821 gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc
     2881 actcgtccga gggcaaagga atagttcgaa ggcctgtcgt gaagcttggg gatcaattct
     2941 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     3001 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3061 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3121 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3181 gctctatggc ttctgaggcg gaaagaacca gctgggccta ctagctactc gggacccctt
     3241 accgaaacat cgccgcattc tgcagaggag tcgagtgtat gtaaacttct gacccactgg
     3301 gaatgtgatg aaagaaataa aagctgaaat gaatcattct ctctactatt attctgatat
     3361 ttcacattct taaaataaag tggtgatcct aactgaccta agacagggaa tttttactag
     3421 gattaaatgt caggaattgt gaaaaagtga gtttaaatgt atttggctaa ggtgtatgta
     3481 aacttccgac ttcaactgta tagggatctg cttcctcgct cactgactcg ctgcgctcgg
     3541 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3601 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3661 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3721 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3781 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3841 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3901 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3961 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4021 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4081 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4141 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4201 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4261 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4321 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4381 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4441 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4501 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4561 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4621 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4681 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4741 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4801 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4861 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4921 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4981 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5041 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5101 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5161 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5221 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5281 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5341 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5401 gggttccgcg cacatttccc cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga
     5461 tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taagatccct ata
//
